PROJECT SUMMARY/ABSTRACT The mission of the WVU Flow Cytometry and Single Cell Core Facility (FCSCCF) is to provide state-of-the-art technologies, instrumentation, and expertise for biomedical research. As part of the Tumor Microenvironment (TME) Center of Biomedical Research Excellence (CoBRE), the WVU FCSCCF has expanded its single cell analysis capabilities with the emplacement of a Fluidigm C1 Single Cell Auto Prep System into the core, along with a Miltenyi gentleMACS Octo Dissociator. This new technology complements the current instrumentation in the facility, which includes two flow cytometers (3-laser BD FACSAria I sorter and 4-laser LSRFortessa analyzer), and a magnetic cell sorter (AutoMACs). A new 4-laser FACSAria III sorter has been purchased with institutional funds and will be installed in early 2017 to replace the existing 3-laser sorter. FACS or MACS-sorted cells prepared following tumor dissociation will be loaded onto a C1 integrated fluidics chip (IFC) for the capture of single cells from complex samples, particularly tumors. RNA (including microRNAs) or DNA will be isolated from the single cells and processed for further downstream applications including RNA/DNA sequencing, or high throughput qRT-PCR. The specific aims of this research core application are to: 1) continue to implement new capabilities into an existing, highly successful core facility in support of research projects for the TME CoBRE and 2) Provide exceptional expertise and training in the area of flow cytometry and single cell analysis to TME CoBRE-funded investigators and other Institutional Development Award (IDeA)-funded projects at WVU. The acquisition of the state-of-the-art Fluidigm C1 system gives WVU FCSCCF users access to cutting edge technology that will significantly advance and enhance their research. The proposal describes the services and training provided by the facility, its management plan, fee structure, quality assurance plan, and strategies to sustain and expand the facility over the life of the project and beyond.